Abstract: Embryo development is a complex and dynamic process that remains poorly understood. Integrated stem cell-based embryo models, termed blastoids, may provide valuable information regarding complex spatiotemporal molecular and cellular processes during embryogenesis ex utero due to their resemblance to natural blastocysts. Moreover, the use of such models avoids the bioethical concerns associated with using natural embryos. Mouse and human cell lines were found capable of self-organizing into blastoids. However, limitations hinder their applications as models for human embryogenesis - mouse blastoids are evolutionary distant, while human blastoids raise bioethical concerns. To bridge this gap, we developed a non-human primate (NHP) blastoid model. Given the close phylogenetic relationship between NHPs and humans, NHP-derived blastoids could closely mimic human embryogenesis while avoiding bioethical concerns surrounding human embryo research. We hypothesized that current induced pluripotent stem cell (iPSC) and blastoid technologies used in human models could be successfully adapted to generate NHP blastoids for comparative developmental studies. This project aimed to generate and characterize NHP blastoids and conduct single-cell RNA-sequencing (scRNA-seq) to create a reference database for comparative studies with human models. In this study, we successfully generated blastoids from North Bornean Orangutan (Pongo pygmaeus pygmaeus). These NHP blastoids exhibited morphology, structure organization, and marker profiles similar to those of human blastoids. Furthermore, downstream assays, including stem cell derivation and in vitro attachment culture, confirmed the functionality of these models, demonstrating their potential as a platform for studying early embryonic development in primates.
